• Applied Biological Chemistry
• /
• v.35 no.3
• /
• pp.165-169
• /
• 1992

Two types of $endo-{\beta}-1,3-glucanases$ were purified from green malt and their basic characteristics were studied. Molecular weights of glucanase I and glucanase II were estimated, by electrophoresis, to be 35,000 and 28,000, respectively. Purified glucanase I and II showed the highest activity at pH $5.0{\sim}7.0$ and $5.0{\sim}8.0$, respectively. The optimal temperature of purified glucanase I and II was $40^{\circ}C$. Purified glucanase I and glucanase II were stable at $40^{\circ}$ for 60 min and at $50^{\circ}$ for 30 min. All enzymes were inactivited by $AgNO_3$ and $HgCl_2$ while those were not activated by various compounds tried. Km values of glucanase I and II were 1.03 mg/ml, 1.20 mg/ml, respectively.

• Microbiology and Biotechnology Letters
• /
• v.19 no.4
• /
• pp.348-355
• /
• 1991

DNA segment encoding $\beta$-1, 4-glucanase of Bacillus subtilis was fused in frame to mouse $\alpha$-amylase signal sequence behind the alcohol dehydrogenase isoenzyme I gene (ADHI) promoter of the yeast expression vector pMS12. To enhance the expression level of the $\beta$glucanase gene in yeast, transcription terminator sequence iso-1-cytochrome c gene (CYCI) was inserted into the recombinant plasmid. The transformants harbouring such recombinant plasmids secreted $\beta$-glucanase into the culture medium. The expresstion level of the $\beta$-glucanase gene was increased about 2-fold caused by inserting the terminator. The amount of the secreted $\beta$-glucanase in culture medium was approximately 60% of the total quantity synthesized.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.40 no.2
• /
• pp.250-254
• /
• 1995

Isolated barley aleurone layers were used to examine response of (1-3)- and $(1-3,1-4)-{\beta}-glucanases{\;}to{\;}GA_3$. Protein content and levels of (1-3)- and $(1-3,1-4)-{\beta}-glucanases$ increased in the presense of added $GA_3$. However, (1-3)- and $(1-3,1-4)-{\beta}-glucanases$ showed different response to $GA_3$ in their production and secretion patterns. $(1-3,1-4)-{\beta}-glucanases$ showed higher increase in enzyme activity than $(1-3)-{\beta}-glucanase$ in the early stage of$GA_3$treatment. Secretion of enzyme by $GA_3$ into the surrounding medium was more enhanced in $(1-3,1-4)-{\beta}-glucanases$ than in $(1-3)-{\beta}-glucanase$, The differential response of the enzymes might be related to the physiological role of the enzymes in germination of barley grain.

• Journal of Life Science
• /
• v.24 no.10
• /
• pp.1046-1054
• /
• 2014

Beta-1,3-glucanase is widely used in various biotechnological and industrial processes, with over-production required to enable versatile utilization. We examined the overexpression of ${\beta}$-1,3-glucanase (EXGA) from Aspergillus oryzae using ${\delta}$-sequence-mediated integration. We constructed $pRS{\delta}$-exgA and $pRS{\delta}K$-exgA plasmids for integration of the EXGA gene into various chromosomes of Saccharomyces cerevisiae. These plasmids contain the ADH1 promoter for constitutive expression, a signal sequence (exoinulinase signal sequence [INU1 s.s]) for secretory production, and a ${\delta}$-sequence for integration of ${\beta}$-1,3-glucanase. The $pRS{\delta}$-exgA plasmid was transformed into the S. cerevisiae $BY4742{\Delta}exg1$ strain, and ${\beta}$-1.3-glucanase was stably overexpressed and secreted. Another plasmid, $pRS{\delta}K$-exgA, was introduced into the S. cerevisiae $BY4742{\Delta}exg1$ (YKY082) strain, and overexpression of ${\beta}$-1,3-glucanase was examined by inducible integration under geneticin selection. The activity of ${\beta}$-1,3-glucanase increased in accordance with a rise in the geneticin concentration, with 0.8 mg/ml of geneticin suitable for overexpression of ${\beta}$-1,3-glucanase. Subsequently, $pRS{\delta}K$-exgA was repeatedly transformed for sequential ${\delta}$-integration. The activity of ${\beta}$-1,3-glucanase reached about 0.063 unit/ml/$OD_{600}$, 0.095 unit/ml/$OD_{600}$, 0.131 unit/ml/$OD_{600}$ and 0.165 unit/ml/$OD_{600}$ by the first, second, third, and fourth round of integration, respectively. According to the increase in the activity of ${\beta}$-1,3-glucanase by sequential ${\delta}$-integration, the copy number (integration rate) of the EXGA gene also increased in various chromosomes. These results suggest that recombinant ${\beta}$-1,3-glucanase activity can be sequentially increased by repeated ${\delta}$-sequence integration.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.39 no.2
• /
• pp.121-127
• /
• 1994

A procedure for the direct detection of (1-3)-$\beta$-glucanase isozymes in electrophoresis gels was developed. The procedure employed the commercial preparation of AZCL-pachyman as a chromogenic substrate for (1-3)-$\beta$-glucanases. The procedure detected the three basic isozymes which have been known to be expressed in germinating barley kernels. A major acidic and a minor isozymes were also detected in germinating kernels. The procedure was proved to be fast, simple and sensitive enough to be used for the analysis of the expression of (1-3)-$\beta$-glucanase isozymes in plant tissues. The detection limit of the procedure for the commercial preparation of Penicillium (1-3)-$\beta$-glucanase was estimated to be as low as 50$\mu$U. The procedure could be used for the investigation of (1-3)-$\beta$-glucanases in laboratories facilitated with ordinary equipments and research personnel.

• KSBB Journal
• /
• v.26 no.5
• /
• pp.427-432
• /
• 2011

In this study, a EXGA gene code for exo-β-1,3-glucanase from Aspergillus oryzae was overexpressed and secretory produced in Saccharomyces cerevisiae. To overexpress the β-1,3-glucanase, pGInu-exgA and pAInu-exgA plasmids having GAL10 and ADH1 promoter, respectively, and exoinulinase signal sequence (Inu s.s) were constructed and introduced in S. cerevisiae SEY2102 and 2805. The recombinant β-1,3-glucanase was successfully expressed and secreted into the medium and the β--1,3-glucanase activity in 2102/pGInu-exgA and 2102/pAInu-exgA strain were 5.01 unit/mL and 4.09 unit/mL, respectively. In the 2805/pGInu-exgA and 2805/pAInu-exgA strain, the β-1,3-glucanase activity showed 3.23 unit/mL and 3.22 unit/mL, respectively. Secretory efficiency in each strain reached 95% to 98%. Subsequently, the recombinant β1,3-glucanase was used for ethanol production. Ethanol productivity in 2102/pAInu-exgA strain was 0.83 g/L when pre-treated Laminaria japonica which has initial reducing sugar of 1.4 g/L was used as substrate. It is assumed that the polysaccharides of Laminaria japonica was effectively saccharified by recombinant β-1,3-glucanase, resulting in increase of ethanol productivity. These results suggested that recombinant β-1,3-glucanase was efficiently overexpressed and secreted in S. cerevisiae SEY2102 as host strain by using ADH1 promoter-Inu s.s system.

• Microbiology and Biotechnology Letters
• /
• v.21 no.3
• /
• pp.263-268
• /
• 1993

Seventeen strains were isolated from soil, cattle rumen, cereal sewage dregs, insect on agar plate containing insoluble glucan as a sole carbon source from immobilized Streptococcus mutans, which produced alpha-1,3 glucanase for lysis of dental plaque. Among these strains isolated from soil, SW-522 and SW-713 that had appeared to produce the high level of alpha-1,3 glucanase, degraded insoluble glucan from S. mutans 97.6% and 49.4%, respectively in 5 hours. The activity of crude alpha-1,3 glucanase from SW-522 was 1.3mg insoluble glucan/min.mg protein. This enzyme was entirely degraded insoluble glucan on glass tube which produced by S. mutans in TH medium with 5% sucrose.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 1992.05a
• /
• pp.20-21
• /
• 1992

• Proceedings of the Korean Society of Crop Science Conference
• /
• 1992.05a
• /
• pp.22-23
• /
• 1992

• Proceedings of the Korean Society of Crop Science Conference
• /
• 1992.05a
• /
• pp.24-25
• /
• 1992